黄瓜抗黑星病相关基因的差异表达分析  

Differential Expression Analysis of Related Resistant Genes to Cladosporium cucumerinum in Cucumber

作　　者：李全辉[1,2] 李锡香[1] 王海平[1] 邱杨[1] 宋江萍[1] 张晓辉[1] 沈镝[1] 

机构地区：[1]中国农业科学院蔬菜花卉研究所,北京100081 [2]青海省农林科学院园艺研究所,西宁810016

出　　处：《植物遗传资源学报》2013年第3期501-506,517,共7页Journal of Plant Genetic Resources

基　　金：国家科技支撑计划项目(2013BAD01B04);农业部园艺作物生物学与种质创制重点实验室资助

摘　　要：以抗黑星病黄瓜材料HX1为试材,接种黑星病菌(Cladosporium cucumerinum)2 h、8 h、20 h、32 h和72 h的叶片作为试验方(Tester),相应的未接种叶片作为对照方(Driver),利用SSH技术,构建了黑星病菌侵染初期的正向和反向cDNA-SSH文库。用巢式引物PCR检测插入片段,获得了200个阳性克隆,通过测序,除去重复序列,共得到105个Unique ESTs,其中50个为singleton,55个为contings。与非冗余蛋白数据库进行BLASTx比对,结果显示,17条ESTs未找到同源序列,88条非重复序列和已知基因的同源性较高,占全部ESTs序列的83.8%,其中86条ESTs与非冗余蛋白数据库已知功能的蛋白具有高度的相似性。结合高密度点阵膜杂交差异筛选,阳性率为75.0%。经初步分析这些序列的功能,差异表达的ESTs功能涉及能量和基础代谢、信号转导、蛋白和核酸代谢、光合作用及逆境中特异表达的基因等方面,为研究黄瓜抗黑星病基因提供了依据。A forward-and reverse-suppression subtractive hybridization libraries were constructed based on a cucumber accession HX1 which showed highly resistant to Cladosporium cucumerinum. The seedling leaves inoculated with Cladosporium cucumerinum on 2 h,8 h、20 h、32 h and 72 h were used as Tester,and that from untreated leave as Driver. Totally 200 SSH eDNA fragments were selected with Nested primer PCR. Through the DNA sequencing and repeated and redundant sequences removing, 105 ESTs including 50 singletons and 55 eontigs were ob- tained. Protein homology search in non-redundant （Nr） protein database revealed that 17 ESTs didn＇t have homologous gene,88 ESTs were highly homologous with known protein, accounting for 83.8% of all EST sequences, of which only 2 ESTs were unknown function protein. The positive rate of these ESTs was 75.0% detected by DIG Nonradioactive Nucleic Acid Labeling and Detection System. The function of these ESTs involved in energy and basic metabolism, signal transduetion, protein and nucleic acid metabolism, photosynthesis, and genes specifically induced under certain stress, etc. This study could provide a basis for further study of the cucumber genes resistance to Cladosporium cucumerinum.

关 键 词：黄瓜 黑星病 抑制性消减杂交 差异表达 

分 类 号：S642.2[农业科学—蔬菜学]